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Constitutive phosphorylation of inhibitor-1 at Ser67 and Thr75 depresses calcium cycling in cardiomyocytes and leads to remodeling upon aging

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Abstract

The activity of protein phosphatase-1 (PP1) inhibitor-1 (I-1) is antithetically modulated by the cAMP-protein kinase A (PKA) and Ca2+-protein kinase C (PKC) signaling axes. β-adrenergic (β-AR) stimulation results in PKA-phosphorylation of I-1 at threonine 35 (Thr35) and depressed PP1 activity, while PKC phosphorylation at serine 67 (Ser67) and/or Thr75 increases PP1 activity. In heart failure, pThr35 is decreased while pSer67 and pThr75 are elevated. However, the role of Ser67/Thr75 phosphorylation in vivo and its effects on Ca2+-cycling are not known. Thus, our aim was to investigate the functional significance of Ser67 and Thr75 phosphorylation in intact hearts. We generated transgenic mice (TG) with cardiac-specific overexpression of constitutively phosphorylated I-1 at Ser67 and Thr75 (S67D/T75D) and evaluated cardiac function. The S67D/T75D cardiomyocytes exhibited significantly depressed Ca2+-kinetics and contractile parameters, compared with wild-type (WT) cells. The decreased Ca2+-cycling was associated with a 27 % increase in PP1 activity, no alterations in PP2 activity and impaired phosphorylation of myosin-binding protein-C (MyBPC). Upon aging, there was cardiac remodeling associated with increases in systolic and diastolic left ventricular internal diameter dimensions (at 16 months), compared with WTs. The results indicate that phosphorylation of I-1 at Ser67 and Thr75 is associated with increased PP1 activity and depressed cardiomyocyte Ca2+-cycling, which manifests in geometrical alterations over the long term. Thus, hyperphosphorylation of these sites in failing hearts may contribute to deteriorative remodeling.

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Acknowledgments

We thank Dr. Wen Zhao for advice on isolation of myocytes, Ms. Andrea Collins for help with the Western Blot technique and Ms. Valerie Lasko for technical expertise in performing mouse catheterization. This work was supported by National Institutes of Health Grants: HL064018 and HL26057.

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Authors and Affiliations

  1. Department of Pharmacology and Cell Biophysics, College of Medicine, University of Cincinnati, 231 Albert Sabin Way, Cincinnati, OH, 45267, USA

    Stela Florea, Wen-Feng Cai, Jiang Qian, Sarah Figueria, Kobra Haghighi & Evangelia G. Kranias

  2. Internal Medicine, Division of Cardiovascular Diseases, College of Medicine, University of Cincinnati, Cincinnati, OH, 45267, USA

    Ahmad Anjak & Jack Rubinstein

  3. Molecular Biology Division, Center for Basic Research, Biomedical Research Foundation of the Academy of Athens, Athens, Greece

    Elizabeth Vafiadaki & Evangelia G. Kranias

  4. Department of Molecular and Cellular Physiology, College of Medicine, University of Cincinnati, Cincinnati, OH, 45267, USA

    John Lorenz

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  1. Stela Florea
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Correspondence to Evangelia G. Kranias.

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Kranias E.G. is a scientific founder of Nanocor.

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Florea, S., Anjak, A., Cai, WF. et al. Constitutive phosphorylation of inhibitor-1 at Ser67 and Thr75 depresses calcium cycling in cardiomyocytes and leads to remodeling upon aging. Basic Res Cardiol 107, 279 (2012). https://doi.org/10.1007/s00395-012-0279-z

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